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Characterization of the complete chloroplast genome of Tilia taishanensis S. B. Liang (Tiliaceae).

ABSTRACT: The complete chloroplast genome of an endangered endemic species in China Tilia taishanensis was sequenced with Illumina HiSeq 2000 platform. It was a typical quadruple structure as other plants of Tilia with 162,803 bp in length, including a large single copy (LSC: 91,114 bp) region and a small single copy (SSC: 20,379 bp) which were separated by a pair of inverted repeats (IRa, b: 25,655 bp) region. The overall GC content is 36.5%. A total of 129 genes was annotated which contained 84 protein-coding genes, 37 tRNA genes, and 8 rRNA genes. ML Phylogenetic analysis compared with 33 expressed chloroplast genomes revealed that T. taishanensis was a sister to other Tilia species.

SUBMITTER: Lu YZ

PROVIDER: S-EPMC7781993 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:Erigeron breviscapus is a famous medicinal plant. However, the limited chloroplast genome information of E. breviscapus, especially for the chloroplast DNA sequence resources, has hindered the study of E. breviscapus chloroplast genome transformation. Here, the complete chloroplast (cp) genome of E. breviscapus was reported. This genome was 152,164bp in length, included 37.2% GC content and was structurally arranged into two 24,699bp inverted repeats (IRs) and two single-copy areas. The sizes of the large single-copy region and the small single-copy region were 84,657 and 18,109bp, respectively. The E. breviscapus cp genome consisted of 127 coding genes, including 83 protein coding genes, 36 transfer RNA (tRNA) genes, and eight ribosomal RNA (rRNA) genes. For those genes, 95 genes were single copy genes and 16 genes were duplicated in two inverted regions with seven tRNAs, four rRNAs, and five protein coding genes. Then, genomic DNA of E. breviscapus was used as a template, and the endogenous 5' and 3' flanking sequences of the trnI gene and trnA gene were selected as homologous recombinant fragments in vector construction and cloned through PCR. The endogenous 5' flanking sequences of the psbA gene and rrn16S gene, the endogenous 3' flanking sequences of the psbA gene, rbcL gene, and rps16 gene and one sequence element from the psbN-psbH chloroplast operon were cloned, and certain chloroplast regulatory elements were identified. Two homologous recombination fragments and all of these elements were constructed into the cloning vector pBluescript SK (+) to yield a series of chloroplast expression vectors, which harbored the reporter gene EGFP and the selectable marker aadA gene. After identification, the chloroplast expression vectors were transformed into Escherichia coli and the function of predicted regulatory elements was confirmed by a spectinomycin resistance test and fluorescence intensity measurement. The results indicated that aadA gene and EGFP gene were efficiently expressed under the regulation of predicted regulatory elements and the chloroplast expression vector had been successfully constructed, thereby providing a solid foundation for establishing subsequent E. breviscapus chloroplast transformation system and genetic improvement of E. breviscapus.

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Characterization of the complete chloroplast genome of Tilia taishanensis S. B. Liang (Tiliaceae).

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